CN110521595A - A method of promoting stem elongation in oriental cherry in vitro culture - Google Patents
A method of promoting stem elongation in oriental cherry in vitro culture Download PDFInfo
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- CN110521595A CN110521595A CN201910828691.7A CN201910828691A CN110521595A CN 110521595 A CN110521595 A CN 110521595A CN 201910828691 A CN201910828691 A CN 201910828691A CN 110521595 A CN110521595 A CN 110521595A
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- oriental cherry
- vitro culture
- stem
- explant
- stem elongation
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
Abstract
The invention discloses a kind of methods of stem elongation in promotion oriental cherry in vitro culture, belong to field of plant tissue culture technique.A kind of method promoting stem elongation in oriental cherry in vitro culture disclosed by the invention, by carrying out disinfection to explant, being cultivated under cold treatment and feux rouges；Overcome the problem of oriental cherry seedling plant strain growth in tissue culture procedures is short and small, elongation of stem is obstructed；The suspend mode for breaking bud using low temperature promotes Stem nematode using hormone and red light irradiation, and stem section length reaches 4.23cm.
Description
Technical field
The present invention relates to field of plant tissue culture technique, in more particularly to a kind of promotion oriental cherry in vitro culture
The method of stem elongation.
Background technique
Oriental cherry (Cerasussp.) is under the jurisdiction of rosaceae, Li Yake, cherry category, sets high 15-25m.Bark Chestnut, it is smooth；
Leaf is oval to ovate-elliptic, and end of blade shape of tail, edge tool is sharp single or weight sawtooth, the short thorn aristiform of increment, two sides are hairless.Grey color or
Pale pink, single-lobe, 3-5 at short umbrella room raceme, British plain spirits, calyx tube mitriform is hairless.Drupe is spherical, and first red then purpling is brown
Color.In April at florescence, floral leaf is same to put.Oriental cherry likes sunlight, deep fertile and well-drained soil is liked, to sodium hydroxide, carbon dioxide
Equal pernicious gases have stronger adsorption capacity, therefore can be used to purify air.Main mutation has: the white oriental cherry of polyphyll, F. pendula Bean,
Magnificent oriental cherry etc. is world-renowned ornamental woody flower, and tree-like grace, spring full-blown flowers are competing to be put, slim and graceful delicate and charming, suitable group in blocks
Plant, be used for Landscape or shade tree, can be used for the fields such as food, drinks, pharmacy, health care product, daily-use chemical industry, have compared with
High ornamental values and the economic values.
Flowering Cherry Cultivars share more than 150 kinds in the whole world, are mainly distributed on Europe, the warmly band of North America, Asia in the Northern Hemisphere
China, Japan and the Korea in continent.Wherein China has about 50 kinds of oriental cherry or so, is distributed mainly on the Yangtze river basin, western part, southwest ground
Area.The breeding of traditional oriental cherry mainly has seed sowing, grafting and cuttage etc..Seminal propagation is although efficient and convenient, but certainly
It is not easy to germinate under the conditions of so, while it is by acquiring, store, modulate the multiple factors such as time and germination percentage and is influenced, current grafting
It is the most common propagation method of oriental cherry, but uses cuttage due to can not find suitable stock to be difficult to carry out the factorial production
Oriental cherry is difficult to take root.Not only have that breeding coefficient is big, the seedling period is short, keeps the steady of excellent inhereditary feature by tissue culture technique
It is qualitative, and a large amount of high quality seedlings can be obtained in a short time.With the maturation of tissue culture technique, more and more scholars with
And scientific research personnel is carrying out in-depth study to it.Mainly there is the culture of the organs such as embryo culture, bud differentiation culture, blade, but
The shorter stem section with oriental cherry of oriental cherry tissue-cultured seedling is difficult to elongation growth.Therefore it provides stem extends in a kind of promotion oriental cherry in vitro culture
Method the problem of being those skilled in the art's urgent need to resolve.
Summary of the invention
In view of this, the present invention provides a kind of methods of stem elongation in promotion oriental cherry in vitro culture.
To achieve the goals above, the present invention adopts the following technical scheme:
A method of promoting stem elongation in oriental cherry in vitro culture, the specific steps are as follows:
(1) 4-6 month acquires explant；
(2) explant is placed in the beaker sterilized, seals mouth with gauze, rinse 1.5-2h under tap water；With 5%
Sodium hypochlorite impregnate 15min, aseptic water washing 2-3 times；+ 0.2% captan of 0.2% benomyl impregnates 10min, sterile water punching
Wash clean, it is spare；
(3) explant after disinfection is seeded in WPM+0.2mgL-16-BA+0.1mg·L-1On NAA culture medium, In
Cold treatment culture 42d under the conditions of 5.5 DEG C；Using red light irradiation, intensity of illumination is 20 μm of ol/ (m2S), light application time 12h/
d；
(4) after cold treatment, continue to cultivate 28d under the conditions of tissue-cultured seedling is transferred to 21-25 DEG C, using red light irradiation,
Intensity of illumination is 20 μm of ol/ (m2S), light application time 12h/d.
Further, the explant is selected from that stalwartness, no disease and pests harm, axillary bud be full, the good branch of growing state.
Further, the explant is the 2-3cm stem section with 1-2 axillary bud.
Further, the pH of step (3) described culture medium is 5.8-6.0.
It can be seen via above technical scheme that compared with prior art, the present disclosure provides a kind of promotion oriental cherry from
The method that stem extends in body culture, overcomes that oriental cherry seedling plant strain growth in tissue culture procedures is short and small, stem extend is obstructed
Problem；The suspend mode for breaking bud using low temperature promotes Stem nematode using hormone and red light irradiation, and stem section length reaches 4.23cm.
Detailed description of the invention
In order to more clearly explain the embodiment of the invention or the technical proposal in the existing technology, to embodiment or will show below
There is attached drawing needed in technical description to be briefly described, it should be apparent that, the accompanying drawings in the following description is only this
The embodiment of invention for those of ordinary skill in the art without creative efforts, can also basis
The attached drawing of offer obtains other attached drawings.
Fig. 1 attached drawing is the aseptic seedling picture that the culture of embodiment 1 obtains；
Fig. 2 attached drawing is the aseptic seedling picture that comparative example culture obtains.
Specific embodiment
Following will be combined with the drawings in the embodiments of the present invention, and technical solution in the embodiment of the present invention carries out clear, complete
Site preparation description, it is clear that described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.It is based on
Embodiment in the present invention, it is obtained by those of ordinary skill in the art without making creative efforts every other
Embodiment shall fall within the protection scope of the present invention.
Oriental cherry maternal plant is selected from Jiangsu Polytechnic College of Agriculture and Forestry's scientific research nursery.
Embodiment 1
A method of promoting stem elongation in oriental cherry in vitro culture, the specific steps are as follows:
(1) fair weather in April is selected, it is full, raw to choose maternal plant stalwartness, no disease and pests harm, axillary bud for 10 AM or so
Long all right annotinous branch, cuts off blade, stem section of the 2-3cm with 1-2 axillary bud is cut into, as explant；
(2) explant is placed in the beaker sterilized, seals mouth with gauze, rinse 1.5-2h under tap water；With 5%
Sodium hypochlorite impregnate 15min, aseptic water washing 2-3 times；+ 0.2% captan of 0.2% benomyl impregnates 10min, sterile water punching
Wash clean, it is spare；
(3) explant after disinfection is seeded in WPM+0.2mgL-16-BA+0.1mg·L-1(pH is NAA culture medium
On 5.8-6.0), cold treatment culture 42d under the conditions of 5.5 DEG C；Using red light irradiation, intensity of illumination is 20 μm of ol/ (m2S),
Light application time is 12h/d；
(4) after cold treatment, continue to cultivate 28d under the conditions of tissue-cultured seedling is transferred to 21-25 DEG C, using red light irradiation,
Intensity of illumination is 20 μm of ol/ (m2S), light application time 12h/d.
Comparative example
A kind of oriental cherry in-vitro culture method, the specific steps are as follows:
(1) fair weather in April is selected, it is full, raw to choose maternal plant stalwartness, no disease and pests harm, axillary bud for 10 AM or so
Long all right annotinous branch, cuts off blade, stem section of the 2-3cm with 1-2 axillary bud is cut into, as explant；
(2) explant is placed in the beaker sterilized, seals mouth with gauze, rinse 1.5-2h under tap water；It rinses
Detergent immersion 10-15min is used after good, it is then clean with aseptic water washing, then with 2000ppm carbendazim+600ppm mould
The mixed solution of plain sodium impregnates 10min, then with aseptic water washing 2-3 times until clean；It is put on superclean bench, prepares to connect
Kind；Every bottle connects 1 material, connects 40 bottles, is repeated 2 times；
(3) explant after disinfection is seeded in WPM+0.5mgL-16-BA+0.1mg·L-1(pH is NAA culture medium
On 5.8-6.0), in 21-25 DEG C of culture 70d, light application time 12h/d, intensity of illumination 2000lx.
After culture, the aseptic seedling obtained to embodiment 1 and comparative example is taken pictures, as a result as shown in Figs. 1-2；And it is right
The stem section length of aseptic seedling measures, and the stem section length average value of 1 aseptic seedling of embodiment is 4.23cm, comparative example aseptic seedling
Stem section length average value is 2.47cm.
The foregoing description of the disclosed embodiments enables those skilled in the art to implement or use the present invention.
Various modifications to these embodiments will be readily apparent to those skilled in the art, as defined herein
General Principle can be realized in other embodiments without departing from the spirit or scope of the present invention.Therefore, of the invention
It is not intended to be limited to the embodiments shown herein, and is to fit to and the principles and novel features disclosed herein phase one
The widest scope of cause.
Claims (4)
1. a kind of method for promoting stem elongation in oriental cherry in vitro culture, which is characterized in that specific step is as follows:
(1) 4-6 month acquires explant；
(2) explant is rinsed into 1.5-2h；15min, aseptic water washing 2-3 times are impregnated with 5% sodium hypochlorite；0.2% benzene comes
Special+0.2% captan impregnates 10min, and aseptic water washing is clean, spare；
(3) explant after disinfection is seeded in WPM+0.2mgL-16-BA+0.1mg·L-1On NAA culture medium, in 5.5 DEG C of items
Cold treatment culture 42d under part；Using red light irradiation, intensity of illumination is 20 μm of ol/ (m2S), light application time 12h/d；
(4) after cold treatment, continue to cultivate 28d under the conditions of tissue-cultured seedling is transferred to 21-25 DEG C, using red light irradiation, illumination
Intensity is 20 μm of ol/ (m2S), light application time 12h/d.
2. a kind of method for promoting stem elongation in oriental cherry in vitro culture according to claim 1, which is characterized in that described outer
Implant is selected from that stalwartness, no disease and pests harm, axillary bud be full, the good branch of growing state.
3. a kind of method for promoting stem elongation in oriental cherry in vitro culture according to claim 1, which is characterized in that described outer
Implant is the 2-3cm stem section with 1-2 axillary bud.
4. a kind of method for promoting stem elongation in oriental cherry in vitro culture according to claim 1, which is characterized in that step
(3) pH of the culture medium is 5.8-6.0.
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